1. Field of the Invention
Although many substances can be assayed by immunoassay, the method depends on the availability of high affinity and specificity antibodies. It is particularly difficult to obtain useful, highly specific antibodies to saccharides, particularly monosaccharides, and other substances, which are naturally present in body fluids, such as NADH, hydroxy acids, steroids, saturated and unsaturated fatty acids, etc. Methods of radioimmunoassay typically require radiolabeled analytes in addition to high affinity and specificity antibodies. Such radiolabeled analytes are frequently difficult or inconvenient to obtain.
Radioenzymatic assays utilize enzymes for the conversion of an analyte to a radiolabeled product. The radiolabel is typically derived from a radioactive enzyme cofactor such as S-adenosyl-L-[methyl-.sup.3 H]methionine, [.sup.14 C]ATP, [.sup.3 H]acetyl CoA, [.sup.14 C]acetyl CoA, [.sup.14 C]adenine, [.gamma.=.sup.32 P]ATP, or [.sup.14 C]CTP. These assays frequently rely on (1) the transfer of a large radiolabeled group such as methyl, acetyl, nucleotide/nucleoside, or (2) phosphorylation.
A method of assay which eliminates the need for high affinity and specificity antibodies and/or radiolabeled analytes would be advantageous. The present invention relates to methods of assay which eliminate the need for both antibodies and radiolabeled analytes.
2. Brief Description of the Related Art
Preparation of isotopically labeled alcohols by exchange of hydrogen atoms for deuterium or tritium atoms has been reported. For example, Roth, C., German Patent No. 2062582 (1972) reports the preparation of stereospecific hydrogen isotope labeled alcohols by use of AND(P) dependent dehydrogenases.
Preparation of isotopically labeled NADH or NADPH has been reported. For example, Valera, V.; et al. Biochem. Biophy. Res. Commun. 1987, 148(1), 515-520 reports the preparation of 4R- and 4S-tritium labeled NADH and NADPH by use of glucose dehydrogenase from Bacillus sp., alcohol dehydrogenase from Thermoanaerobium brockii and D-[1-.sup.3 H]glucose.
Enzymatic incorporation of a radiochemical label into an enzyme substrate or removal of a label from a substrate by enzyme catalysis has reportedly been used to study enzyme mechanisms. Further, microbial conversion of radiolabeled nutrients to a radioactive product, such as [.sup.14 C]CO.sub.2, has reportedly been used to detect microorganisms.
Radioenzymatic assays involving methyl, acetyl, nucleotide, nucleoside, and phosphate group transfers have been reported.
Hussain, M. N.; et al. Clin. Chem. 1985, 31(11), 1861-1864 reports the use of catechol-O-methyltransferase (COMT) and S-adenosyl-L-[methyl-.sup.3 H]methionine ([.sup.3 H]SAM) in the analysis of dopamine, norepinephrine, and epinephrine.
Nagel-Hiemke, M.; et al. J. Biochem. Biophvs. Meth. 1981, 4(5-6), 261-270 reports the use of COMT and [.sup.3 H]SAM in the analysis of dihydroxymandelic acid, dihydroxyphenylglycol, and dihydroxyphenylacetic acid.
Johnson, G. A.; et al. U.S. Pat. No. 4,288,542 (1981) reports the use of COMT and [.sup.3 H]SAM in the analysis of catecholamines.
Vlachakis, N. D. U.S. Pat. No. 4,311,790 (1982) reports the use of COMT and [.sup.3 H]SAM in the analysis of 3,4-dihydroxyphenylglycol.
Johnson, G. A.; et al. German Patent No. 2717306 (1977) and equivalent U.S. Pat. Nos. 4,242,222 (1980), 4,242,456 (1980), 4,284,587 (1981), and 4,287,368 (198) reports the use of COMT and [.sup.3 H]SAM in the analysis of epinephrine, norepinephrine, and dopamine.
Hussain, M. N.; et al. Anal. Biochem. 1981, 111(1), 105-110 reports the use of hydroxyindole-O-transferase (HIOMT) and ([.sup.3 H]SAM) in the analysis of indolealkylamines such as serotonin and acetylserotonin.
Giulidori, P.; et al. Anal. Biochem. 1984, 137(1), 217-220 reports the use of HIOMT and [.sup.3 H]SAM in the analysis of S-adenosyl-L-methionine (SAM).
Harvima, R. J.; et al. Clin. Chim. Acta 1988, 171(2-3), 247-256 and Henry, D. P.; et al. U.S. Pat. No. 4,769,322 (1988) describe the use of histamine-N-methyl- transferase (HNMT) and [.sup.3 H]SAM in the analysis of histamine.
Bowsher, R. R.; et al. U.S. Pat. No. 4,649,1077 (1987) reports the use of phenylethanolamine-N-methyl transferase (PNMT) and [.sup.3 H]SAM in the analysis of noradrenalin.
Vlachakis, N. D. U.S. Pat. No. 4,275,150 (1981) and De Quattro, V.; et al. U.S. Pat. No. 4,591,551 (1986) report the use of PNMT and [.sup.3 H]SAM in the analysis of normetanephrine and/or octopamine.
Robison, L. R.; et al. Antimicrob. Agents Chemother. 1978, 13(1), 25-29 reports the use of chloramphenicol acetyltransferase (CAT) and [.sup.14 C]acetyl Coenzyme A ([.sup.14 C]acetyl CoA) in the analysis of chloramphenicol.
Stevens, P.; et al. Antimicrob. Agents Chemother. 1975, 7(3), 374-376 reports the use of kanamycin acetyltransferase (KAT) and [.sup.3 H]acetyl Coenzyme A ([.sup.3 H]acetyl CoA) in the analysis of aminoglycosides such as amikacin, tobramycin, and sisomicin.
Chan, A.; et al. Life Sci. 1981, 28(6), 697-703 reports the use of acetyl CoA synthetase/serotonin-N-acetyl transferase (ACS/SNAT) and [.sup.3 H]acetate/tryptamine in the analysis of Coenzyme A.
Consolo, S.; et al. J. Neurochem. 1987, 48(5), 1459-1465 reports the use of acetyl-CoA:choline-O-acetyltransferase (ACCOAT) and [.sup.3 H]acetyl-CoA in the analysis of acetylcholine.
Kredich, N. M.; et al. Anal. Biochem. 1981, 116(2), 503-510 reports the use of S-adenosyl-L-homocysteine hydrolase (AHH) and [.sup.3 H]adenosine in the analysis of S-adenosyl-L-homocysteine and L-homocysteine.
Smith, D. H.; et al. N. Engl. J. Med. 1972, 286(11), 583-586 reports the use of gentamicin adenyl transferase (GAT) and [.sup.14 C]ATP in the analysis of aminoglycosides such as gentamicin.
Ipata, P. L.; et al. Anal. Biochem. 1987, 164(2), 411-417 reports the use of adenine phosphoribosyltransferase (APT) and [.sup.14 C]adenine in the analysis of 5-phosphorylribosyl-1-pyrophosphate.
Brackmann, Th.; et al. Anal. Biochem. 1983, 130(2), 369-375 reports the use of CMP-N-acetylneuraminic acid synthetase (CNAAS) and [.sup.14 C]CTP in the analysis of N-acetylsialic acid and CMP-N-acetylsialic acid.
Schneider, P. B. J. Lioid Res. 1977, 18(3), 396-399 reports the use of glycerol kinase (GK) and [.gamma.-.sup.32 P]ATP in the analysis of glycerol and acylglycerol.
The purification and properties of Klebsiella aeroegenes D-arabinitol dehydrogenase has been reported by Neuberger, et al., in Biochem. J., 183 (1979) 31-42.